Aminoglycoside antibiotics

ABSTRACT

6-0- AND 3&#39;-0-D-glycosyl analogs of 4-0-(α-D-glycosyl)-2-deoxystreptamine, 6-0- and 3&#39;-0-D-glycosyl ortho esters of 4-0-(α-D-glycosyl)-2-deoxystreptamine, novel aminoglycoside antibiotics, and novel intermediates are prepared by a new chemical process. The compounds have utility as antibacterial agents or as intermediates to make antibacterially-active compounds.

BACKGROUND OF THE INVENTION

Microbially produced aminoglycosides, possessing the 2-deoxystreptaminemoiety, have either a pentofuranosyl substituent at the 5-O position ora hexopyranosyl substituent at the 6-O position. Examples of antibioticswith a pentofuranosyl substituent at the 5-O position are paromomycin,neomycin, lividomycin and ribostamycin. Examples of antibiotics having ahexopyranosyl substituent at the 6-O position are kanamycin B andgentamicin C_(1a). Subsequent to the subject invention, a publication byT. Ogawa, T. Takamoto and S. Hanessian, Tetrahedron Letters, 46, 4013(1974), discloses the preparation of a 6-O-pentofuranosylparomamineanalog. Paromamine differs from neamine by having a hydroxyl at the 6'position instead of an amino moiety.

BRIEF SUMMARY OF THE INVENTION

The subject invention concerns novel aminoglycosides, nontoxicpharmaceutically acceptable acid addition salts thereof, intermediates,and a novel process for their preparation. More particularly, thesubject invention concerns 6-O- and 3'-O-D-glycosyl analogs of4-O-(α-D-glycosyl)-2-deoxystreptamine, novel aminoglycoside antibiotics,novel intermediates, and a chemical process for the preparation thereof.The process comprises essentially six steps: (1) the aminogroups of thestarting aglycone, are blocked; (2) the 5,6-hydroxyls are blocked as theketal; (3) the 3',4'-hydroxyls are acylated; (4) the ketal group isremoved; (5) the desired sugar moiety is added using a variation of thewell-known Koenigs-Knorr glycosylation reaction; and, (6) the blockinggroups are removed to afford the new aminoglycoside antibiotic. When the3'-O-D-glycosyl analog is made, Step 3 and Step 4 are omitted.

The subject invention process is novel and advantageous for thepreparation of the new aminoglycosides for several reasons. The blockingof the amino groups as the trifluoroacetates departs from the usualchemical approach using the carbobenzyloxy group as a blocking agent.The trifluoroacetyl group has advantages over the carbobenzyloxy groupin its preparation; it is easier to remove (mild alkali vs.hydrogenolysis); it imparts much greater solubility in organic solventsthus permitting easier manipulation and purification (for example, withchromatography); and, since it is more volatile, intermediates can beexamined by vapor phase chromatography, thus facilitating the synthesis.Further, the very selective formation of a ketal is surprising andunexpected in view of prior work in the art. For example, Umezawa, etal., Bull Chem. Soc. Japan, 42, 537 (1969), when working withcarbobenzyloxy-blocked neamine, formed a difficultly separable mixtureof monoketals.

The novel six step process, briefly described above, can beschematically depicted as follows showing, for convenience, a specificsugar. However, it is to be understood that other sugars, as disclosedherein, can be used to make novel aminoglycoside antibiotics. Also, forconvenience, neamine is the depicted aglycone. The scope of4-O-(α-D-glycosyl)-2-deoxystreptamine is given infra. ##SPC1## ##SPC2##

Novel ortho esters are prepared by reacting compound 5 with the desiredsugar moiety in the presence of a strong base. This reaction can beschematically depicted as follows: ##SPC3##

The protecting groups on compound 8 are then removed following theprocedures of Step 6, above, to afford the ortho ester. Substitution ofthe sugar at the 3-O- position ortho esters is accomplished by omittingthe acylation step (Step 3) and reacting compound 3 with the desiredsugar moiety in the presence of a strong base. The resulting compound isthen subjected to mild acid hydrolysis to remove the ketal group at the5,6 position. The remaining acyl groups on the sugar moiety are removedby alkaline hydrolysis.

DETAILED DESCRIPTION OF THE INVENTION

Novel aminoglycoside antibiotics, which are 6-O- and 3'-O-glycosylanalogs of 4-O-(α-D-glycosyl)-2-deoxystreptamine and 6-O- and3'-O-glycosyl ortho esters of 4-O-(α-D-glycosyl)-2-deoxystreptamine canbe prepared by the novel process described herein.

The following 4-O-(α-D-glycosyl)-2-deoxystreptamines can be used as thestarting material aglycone in addition to neamine, disclosed above:##SPC4## ##SPC5##

The subject invention process yields novel 6-O- and 3'-O-D-glycosylanalogs of neamine and paromamine. Since the remaining4-O-(α-D-glycosyl)-2-deoxystreptamines, as disclosed above, eithercontain 3 hydroxyls in the B ring (left-hand ring) or lack a C-3hydroxyl, only novel 6-O-D-glycosyl analogs of these aglycones are made.

The starting material aglycones are well known in the art and can beprepared by the procedures given in the following publications:

Neamine (Neomycin A): Prep. from neomycin B, U.S. 2,691,675. Synthesisfrom paromine, S. Umezawa and K. Tatsuta, Bull. Chem. Soc. Japan, 40,2371-75 (1967).

Paromamine: S. Umezawa and S. Koto, Bull. Chem. Soc. Japan, 39, 2014(1966).

Kanamine: S. Umezawa, T. Tsuchiya, J. Antib., 15, 51-52 (1962).

Nebramine (Tobramine): K. F. Koch, J. A. Rhoades, E. W. Hagaman and E.Wenkert, J. Amer. Chem. Soc., 96, 3300-05 (1974). [however havetypographical error between neamine and nebramine]

Lividamine: T. Oda, T. Mori and Y. Kyotani, J. Antib., 24, 503-10(1971).

Gentamine C_(1a), C₁ and C₂ : D. J. Cooper, P. J. L. Daniels, M. D.Yudis, H. M. Marigliano and (in part) R. D. Gulkrie and S. T. K. Bukari,J. Chem. Soc. (C), 3126-29 (1971). More data J. B. Morton, R. C. Long,P. J. L. Daniels, R. W. Tkach and J. H. Goldstein, J. Amer. Chem. Soc.,95, 7464-69 (1973).

Gentamine C_(2b) : P. J. L. Daniels, C. Luce, T. L. Nagathusushan, R. S.Faut, D. Schumacher, H. Kermann and J. Ilavasky, J. Antib., 28, 35(1975).

R. S. Egan, R. L. DeVault, S. L. Mueller, M. I. Levenberg, A. C.Sinclair and R. S. Stanasyck, J. Antib., 28, 29-34 (1975).

Gentamine B₁ : Belg. Pat. No. 768,796, June 21, 1971.

In the novel process, the amino groups in the aglycone starting materialare first blocked by a suitable blocking group. The preferred blockinggroup is trifluoroacetyl. Thus, neamine (1), in the preferred embodimentof this process step, hereinafter referred to as Step 1 for convenience,is reacted, as a suspension in acetonitrile, with trifluoroaceticanhydride in the presence of an organic base, for example,triethylamine. The trifluoroacetic anhydride is added to the neaminesuspension at 15° ± 5° over a period of 30 minutes. The reaction mixtureis stirred at ambient temperature for about one hour and the solvent isthen evaporated in vacuo. The desired product (2) is then recovered bysolvent extraction with ethyl acetate and crystallization from ethanol.

Suitable substitutions for trifluoroacetic anhydride in Step 1 arepentafluoropropionic anhydride and the ethylthio ester oftrifluoroacetic acid, S-ethyl trifluorothioacetate. A suitablereplacement for acetonitrile in Step 1 is ethyl acetate or othersolvents in which the reaction products are soluble. The reaction can becarried on over a temperature range of 0° to the boiling point of thereactants. Those skilled in the art recognize that the lower thereaction temperature, the longer the reaction time. The recovery of theproduct from Step 1 is accomplished by well-known art procedures. Inplace of ethyl acetate, which is preferred as the extraction solvent,other water-insoluble solvents, for example butyl acetate, and the like,can be used.

Step 2 of the invention process is concerned with forming the 5,6-ketalof the compound obtained in Step 1. In a preferred Step 2 process,product (2) in acetonitrile and 2,2-dimethoxypropane containingtrifluoroacetic acid is heated at reflux for about 3/4 of an hour.Thereafter, the basic resin IRA-45 (OH⁻) is added to the reactionsolution to remove the acid catalyst. Monoketal compound (3) isrecovered from the reaction by well-known chromatographic techniques.

The 2,2-dimethoxypropane of Step 2 can be replaced by other dialkoxylower alkanes where in the alkoxy and lower alkane can be from 1 to 8carbon atoms, inclusive. Preferably, the lower alkane radicals are thesame, but they can be different. Examples of suitable dialkoxy loweralkanes are 2,2-diethoxypropane, 2,2-dipropoxypropane,2,2-dibutoxypropane, 2,2-dipentoxypropane, 2,2-dihexoxypropane,2,2-diheptoxypropane, 2,2-dioctoxypropane, dimethoxymethane,3,3-dimethoxypentane, 4,4-dimethoxyheptane, and the like, and2-ethoxy-2-methoxypropane, and the like.

The acid catalyst trifluoroacetic acid in Step 2 can be replaced byp-toluenesulfonic acid or strong inorganic acids, for example, HCl, H₂SO₄, and the like. It is desirable to control the amount of acidcatalyst used since an excess amount will result in the formation of thediketal compound instead of the desired monoketal. If the diketal isformed, it can be converted to the monoketal by selective methanolysisprocedures known in the art employing methanol and an acid catalyst, forexample, trifluoroacetic acid.

In the preferred process, the removal of the acid catalyst in Step 2upon completion of the reaction is accomplished by mixing the reactionsolution with Amberlite IRA-45 (OH⁻), a basic resin. Other resins whichcan be used are obtained by chloromethylating by the procedure given onpages 88 and 97 of Kunin, Ion Exchange Resins, 2nd ed. (1958), JohnWiley and Sons, Inc., polystyrene crosslinked, if desired, withdivinylbenzene, prepared by the procedure given on page 84 of Kunin,supra, and quaternizing with trimethylamine or dimethylethanolamine bythe procedure given on page 97 of Kunin, supra. Anion exchange resins ofthis type are marketed under the tradenames Dowex 2, Dowex 20, AmberliteIRA-400 (OH⁻), Amberlite IRA-410 (OH⁻), Amberlite IRA-401 (OH⁻), DuoliteA-102 and Permutit S.1.

The acid catalyst of Step 2 also can be removed by the use of insolublebasic salts, for example, barium carbonate, lead carbonate, and thelike.

The acylation of the 3' and 4'-hydroxyls of Step 3 can be carried out byacylating procedures well-known in the art. The preferred process usesp-nitrobenzoyl chloride as the acylating agent because it gives theacylated product (4) which produces ultraviolet light visibility on thinlayer chromatography during the Koenigs-Knorr glycosylation of Step 5.However, other acylating agents can be used to give the acylatedproduct. The acylation is carried out in the presence of an acid-bindingagent. Suitable acid-binding agents include amines such as pyridine,quinoline, and isoquinoline, and buffer salts such as sodium acetate.The preferred base is pyridine. Carboxylic acids suitable for acylationinclude (a) saturated or unsaturated, straight or branched chainaliphatic carboxylic acids, for example, acetic, propionic, butyric,isobutyric, tert-butylacetic, valeric, isovaleric, caproic, caprylic,decanoic, dodecanoic, lauric, tridecanoic, myristic, pentadecanoic,palmitic, margaric, stearic, acrylic, crotonic, undecylenic, oleic,hexynoic, heptynoic, octynoic acids, and the like; (b) saturated orunsaturated, alicyclic carboxylic acids, for example,cyclobutanecarboxylic acid, cyclopentanecarboxylic acid,cyclopentenecarboxylic acid, methylcyclopentenecarboxylic acid,cyclohexanecarboxylic acid, dimethylcyclohexanecarboxylic acid,dipropylcyclohexanecarboxylic acid, and the like; (c) saturated orunsaturated, alicyclic aliphatic carboxylic acids, for example,cyclopentaneacetic acid, cyclopentanepropionic acid, cyclohexaneaceticacid, cyclohexanebutyric acid, methylcyclohexaneacetic acid, and thelike; (d) aromatic carboxylic acids, for example, benzoic acid, toluicacid, naphthoic acid, ethylbenzoic acid, isobutylbenzoic acid,methylbutylbenzoic acid, and the like; and (e) aromatic aliphaticcarboxylic acids, for example, phenylacetic acid, phenylpropionic acid,phenylvaleric acid, cinnamic acid, phenylpropiolic acid, andnaphthylacetic acid, and the like. Also, suitable halo-, nitro-,hydroxy-, amino-, cyano-, thiocyano-, and lower alkoxyhydrocarboncarboxylic acids include hydrocarboncarboxylic acids as given abovewhich are substituted by one or more of halogen, nitro, hydroxy, amino,cyano, or thiocyano, or loweralkoxy, advantageously loweralkoxy of notmore than six carbon atoms, for example, methoxy, ethoxy, propoxy,butoxy, amyloxy, hexyloxy groups, and isomeric forms thereof. Examplesof such substituted hydrocarbon carboxylic acids are:

mono-, di- and trichloroacetic acid;

α- and β-chloropropionic acid;

α- and γ-bromobutyric acid;

α- and δ-iodovaleric acid;

mevalonic acid;

2- and 4-chlorocyclohexanecarboxylic acid;

shikimic acid;

2-nitro-1-methyl-cyclobutanecarboxylic acid;

1,2,3,4,5,6-hexachlorocyclohexanecarboxylic acid;

3-bromo-2-methylcyclohexanecarboxylic acid;

4- and 5-bromo-2-methylcyclohexanecarboxylic acid;

5- and 6-bromo-2-methylcyclohexanecarboxylic acid;

2,3-dibromo-2-methylcyclohexanecarboxylic acid;

2,5-dibromo-2methylcyclohexanecarboxylic acid;

4,5-dibromo-2-methylcyclohexanecarboxylic acid;

5,6-dibromo-2-methylcyclohexanecarboxylic acid;

3-bromo-3-methylcyclohexanecarboxylic acid;

6-bromo-3-methylcyclohexanecarboxylic acid;

1,6-dibromo-3-methylcyclohexanecarboxylic acid;

2-bromo-4-methylcyclohexanecarboxylic acid;

1,2-dibromo-4-methylcyclohexanecarboxylic acid;

3-bromo-2,2,3-trimethylcyclopentanecarboxylic acid;

1-bromo-3,5-dimethylcyclohexanecarboxylic acid;

homogentisic acid, o-, m-, and p-chlorobenzoic acid;

anisic acid;

salicyclic acid;

p-hydroxybenzoic acid;

β-resorcylic acid;

gallic acid;

veratric acid;

trimethoxybenzoic acid;

trimethoxycinnamic acid;

4,4'-dichlorobenzilic acid;

o-, m-, p-nitrobenzoic acid;

cyanoacetic acid;

3,4- and 3,5-dinitrobenzoic acid;

2,4,6-trinitrobenzoic acid;

thiocyanoacetic acid;

cyanopropionic acid;

lactic acid;

ethoxyformic acid (ethyl hydrogen carbonate); and the like.

Product (4), obtained in Step 3, is subjected to mild acid hydrolysis toremove the ketal group. In the preferred process of Step 4, a solutionof (4) in about a 66% acetic acid solution is warmed at 65° for about 4hours. Recovery of the desired product (5) is accomplished by removal ofthe solvent and use of standard chromatographic procedures.

The acetic acid of Step 4 can be replaced by other mild acids, forexample, propionic and oxalic, which will not cause ester hydrolysis. Ifa stronger acid is used, for example, trifluoroacetic acid, hydrochloricsulfuric or trichloroacetic acid, then the reaction time will be shorterand the temperature of the reaction lower. The temperature of thereaction can be varied over a range of about 10° to about 100° dependingon the acid used.

The glycosylation of product (5) is performed, according to Step 5, byuse of the well-known Koenigs-Knorr reaction. It is critical forsuccessful yields in Step 5 that the reaction be conducted underanhydrous conditions. In the preferred process of Step 5, the anhydrousconditions are obtained by distillation of benzene from the reactionmixture and use of an atmosphere of dry N₂. In the preferred process ofStep 5, a solution of desired sugar moiety in nitromethane-benzene, asthe bromide or chloride, and with the hydroxyls blocked by acetylgroups, is reacted with compound (5) in the presence of Hg(CN)₂ underreflux to give compound (6).

Alternatively, the hydroxyls of the sugar moiety of Step 5 can beblocked by benzyl groups to give the benzyl ether and these benzylgroups can subsequently be removed by hydrogenolysis:

Nitromethane is the preferred solvent in Step 5 because mercuric cyanideis relatively soluble in this solvent. Other solvents which can be usedare ethyl acetate, acetonitrile and dimethylformamide since mercuriccyanide is somewhat soluble in these solvents.

Mercuric cyanide of Step 5 can be replaced by other mercury salts, forexample, mercuric bromide, mercuric chloride, mercuric oxide, and thelike. Further, silver salts, for example, silver carbonate, silverperchlorate, and the like, can be used in place of mercuric cyanide.

The temperature of the reaction in Step 5 can range from about roomtemperature to about the boiling point of the solvent used.

An excess of the sugar bromide or chloride is necessary in Step 5 inorder to complete the reaction. Thus, the ratio of sugar bromide orchloride to compound (5) should be at least 2:1 and possibly as high as10:1. A great excess is not desirable since recovery problems will begreater.

Step 6 of the invention process is conducted to simultaneously hydrolyzethe esters and amino protecting groups. In a preferred embodiment ofStep 6, a solution of compound (6) and 2N NaOH in methanol is heated atreflux for about 15 minutes. The methanol is then removed in vacuo,water is added to the solution, and compound (7) is recovered bysubjecting the solution to standard ion exchange procedures. Any strongaqueous alkali, for example, potassium hydroxide, barium hydroxide,ammonium hydroxide, and the like, can be substituted for the sodiumhydroxide. The aqueous alkali can be more dilute than 2N but,advantageously, not much stronger so that the protective groups areselectively removed.

The process for preparing ortho esters proceeds from compound (5) tocompound (8). In a preferred embodiment of this process, a solution of(5) in triethylamine is reacted with the desired sugar halide at refluxto give compound (8). Any strong organic base, for example,1,4-bis-(dimethylamino)-naphthalene, can be used instead oftriethylamine. The temperature of the reaction can range from 0° -reflux. The protecting groups on compound (8) can be removed followingthe procedure of Step 6 to afford the desired ortho ester (9). However,care must be taken as the ortho ester is somewhat hydrolyzed by alkalithough at a lesser rate than the other groups.

3'O-D-glycosyl ortho esters are prepared in like manner starting withcompound (3). The ketal and protective groups are removed as disclosedabove.

Novel 1-N-AHBA derivatives of the compounds of the subject invention canbe formed by use of processes well known in the art. These derivativespotentiate the antibacterial activity and make the antibiotic moreresistant to enzymatic inactivation. Thus, these derivatives can be usedfor the same antibacterial purposes as the parent compounds.

The novel 1-N-AHBA derivatives of the subject invention can be made ofany compound of the invention by reacting such compound with one whichcontains three to five carbon atoms, has an α-hydroxyl group in theL-configuration, and has an ω-aminogroup. These compounds can be shownas follows wherein neamine, for convenience, is shown as the aglycone:##SPC6##

wherein X and Y = H, and substituted glycosyl, except that X and Y arenot the same. n is an integer of from 0 to 2, inclusive.

The preparation of the above 1-N-AHBA derivatives can proceed by firstblocking the 6'-amino. This can be accomplished by the reaction of theaminoglycoside with N-benzyloxycarbonyloxylsuccinimide in aqueousdimethylformamide. The 6'-N-carbenzoxyamino glycoside thus formed isselectively 1-N-acylated withL(-)γ-benzyloxycarbonylamino-α-hydroxybutyric acid, N-hydroxysuccinimideester in aqueous ethylene glycol dimethyl ether. The carbobenzoxy groupsat 6'-N and at the γ-N can then be removed hydrogenolysis using, forexample, palladium on charcoal as catalyst. The above procedure isdisclosed in Kawaguchi, Naito, Nakagawa and Fujisawa, J. Antibiotics,25, 695 (1972), and in U.S. Pat. No. 3,781,268.

Other more elegant methods to make 1-N-AHBA derivatives can be used asreviewed by Umezawa in, Adv. Appl. Microbiol., 18, 174 (1973).

Alternatively, the 1-N-AHBA group can be introduced into the startingcompound neamine (1). 1-N-AHBA neamine is disclosed in an article by R.Akita et al., J. Antibiotics, 26, 365 (1973) and Tsukuira, ibid., p.351. If this procedure is used, the α-hydroxy would require blockingwith p-nitrobenzoyl chloride at the same time as the 3'-O and 4'-O esteris formed. The blocking group can then be removed by the procedures ofStep 6, described herein.

Glycosyl halides, as defined by Wolfrom and Szarek, The Carbohydrates,Vol. 1A, p. 239, Pigman and Horton editors, Academic Press, New York(1972), are "saccharide derivatives in which the hydroxyl group of theanomeric center of the aldose or ketose is replaced by a halogen atom."This definition is used herein with the following limitations: (a) 5-8carbon atoms with various configurations of hydroxyl groups; (b)1-chloro or 1-bromo sugars in either the 1-α or 1-β-halo configuration;(c) hydroxy groups blocked as acylates (acetate or benzoate), or asbenzyl ethers; and (d) amino, alkylamino or dialkylamino sugarsincluding, for example, 2-amino-2-deoxy, 3-amino-3-deoxy,4-amino-4-deoxy, 5-amino-5deoxy, 6-amino-6-deoxy,2,6-diamino-2,6-dideoxy, and their N-mono and N,N.sub. 1 -dialkylsubstitutes. A rather complete disclosure of glycosyl halides can befound in the book The Amino Sugars, Vol. 1A, Academic Press, N.Y. (1969)by Jeanloz, and in the publication by L. J. Haynes and F. H. Newth,"Glycosyl Halides And Their Derivatives", Advances in CarbohydrateChemistry, Vol. 10, Academic Press, 1955, pages 247-254.

The generic structure of the glycosyl halides which can be used in thesubject invention can be shown as follows: ##SPC7##

wherein Z is Cl or Br; R₁ = OAc, ##STR1## OCH₂ φ, NHR', NR'-alkyl; alkyl= 1-5 carbon atoms, inclusive; R' = acyl of from 1 to 8 carbon atoms,inclusive; Ac = acetyl; and ##SPC8##

wherein Ac is acetyl.

A sub-generic structure of the glycosyl halides which can be used in thesubject invention to make corresponding aminoglycoside antibiotics canbe shown as follows: ##SPC9##

wherein Z is Cl or Br, and R₁ is OAc or NHAc wherein Ac is acetyl.

A further sub-generic structure of the glycosyl halides which can beused in the subject invention to make corresponding aminoglycosideantibiotics can be shown as follows: ##SPC10##

wherein Z is Cl or Br and Ac is acetyl.

Examples of glycosyls (as halides) of sugars which can be used in thesubject invention are:

2,3,4,6 Tetra-O-acetyl-α-D-altropyranosyl chloride

2,3,4-Tri-O-acetyl-β-L-arabinopyranosyl chloride -O- and -O-(α O

3,4-di-O-acetyl-2-deoxy-D-ribopyranosyl chloride

2,3,4,6-Tetra-O-acetyl-α-D-galactopyranosyl chloride

2,3,4,6 Tetra-O-acetyl-β-D-galactopyranosyl chloride

2,3,4,6-Tetra-O-acetyl-α-D-glucopyranosyl chloride

2,3,4,6-Tetra-O-benzoyl-α-D-glucopyranosyl chloride

2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl chloride

2,3,4,6-Tetra-O-acetyl-α-D-mannopyranosyl chloride

2,3,4,6-Tetra-O-benzoyl-α-D-mannopyranosyl chloride

2,3,4-Tri-O-acetyl-α-L-rhamnopyranosyl chloride

2,3,4-Tri-O-benzoyl-α-L-rhamnopyranosyl chloride

2,3,5-Tri-O-acetyl-α-D-ribofuranosyl chloride

2,3,4-Tri-O-benzoyl-α-D-ribopyranosyl chloride

2,3,4-Tri-O-acetyl-β-D-ribopyranosyl chloride

2,3,4-Tri-O-benzoyl-β-D-ribopyranosyl chloride

2,3,4-Tri-O-acetyl-α-D-xylopyranosyl chloride

2,3,4-Tri-O-acetyl-β-D-xylopyranosyl chloride

2,3,4-Tri-O-acetyl-6-deoxy-α-D-glucopyranosyl chloride

2,3,4-Tri-O-acetyl-β-D-arabinopyranosyl bromide

2,3,4-Tri-O-benzoyl-β-D-arabinopyranosyl bromide

3,4,6-Tri-O-acetyl-2-deoxy-α-D-glucopyranosyl bromide

3,4,6-Tri-O-benzoyl-2-deoxy-α-D-glucopyranosyl bromide

2,3,4-Tri-O-acetyl-6-deoxy-α-D-glucopyranosyl bromide

1,3,4,5-Tetra-O-acetyl-β-D-fructopyranosyl bromide

1,3,4,5-Tetra-O-benzoyl-β-D-fructopyranosyl bromide

2,3,4,6-Tetra-O-acetyl-α-D-galactopyranosyl bromide

2,3,4,6-Tetra-O-acetyl-α-D-glucopyranosyl bromide

2,3,4-Tri-O-acetyl-6-O-benzoyl-α-D-glucopyranosyl bromide

2,3,4-Tri-O-acetyl-6-O-methyl-α-D-glucopyranosyl bromide

6-O-Acetyl-2,3,4-tri-O-benzyl-α-D-glucopyranosyl bromide

2,3,4,6-Tetra-O-acetyl-α-D-mannopyranosyl bromide

2,3,4,6-Tetra-O-benzoyl-α-D-mannopyranosyl bromide

2,3,4-Tri-O-acetyl-α-L-rhamnopyranosyl bromide

2,3,4-Tri-O-benzoyl-α-L-rhamnopyranosyl bromide

2,3,4-Tri-O-acetyl-β-D-ribopyranosyl bromide

2,3,4-Tri-O-benzoyl-β-D-ribopyranosyl bromide

2,3,4-Tri-O-benzoyl-D-xylopyranosyl bromide

2,3,4-Tri-O-acetyl-L-xylopyranoxyl bromide

2,3,4-Tri-O-benzoyl-L-xylopyranoxyl bromide

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-α-D-glucopyranosyl bromide

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-α-D-glucopyranosyl chloride

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl chloride

2-Benzamido-3,4,6-tri-O-benzoyl-2-deoxy-α-D-glucopyranosyl bromide

3,4,6-Tri-O-acetyl-2-benzamido-2-deoxy-α-D-glucopyranosyl chloride

3,4,6-Tri-O-acetyl-2-[(benzyloxycarbonyl)-amino]-2-deoxy-α-D-glucopyranosylbromide

3,4,6-Tri-O-acetyl-2-deoxy-2-(2,4-dinitroanilino)-α-D-glucopyranosylbromide

2-Acetamido-3,4-di-O-acetyl-2-deoxy-D-ribofuranosyl chloride

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-D-galactopyranosyl bromide

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-α-D-galactopyranosyl chloride

3-Acetamido-2,4,6-tri-O-acetyl-3-deoxy-α-D-mannopyranosyl bromide

3-Acetamido-2,4,6-tri-O-acetyl-3-deoxy-α-D-mannopyranosyl chloride

2,4,6-Tri-O-acetyl-3-[(benzyloxycarbonyl)-amino]-3-deoxy-α-D-glucopyranosylbromide

2,3,4-Tri-O-acetyl-6-[(benzyloxycarbonyl)-amino]-6-deoxy-α-D-glucopyranosylbromide

2,4,6-Tri-O-acetyl-3-[(benzyloxycarbonyl)-amino]-3-deoxy-D-glucopyranosylchloride

2,3,4-Tri-O-acetyl-6-[(benzyloxycarbonyl)-amino]-6-deoxy-D-glucopyranosylchloride

The above are known and available glycosyl halides as disclosed in D.Horton, Monosaccharide Amino Sugars, in "The Amino Sugars", Vol. 1A,Editor R. W. Jeanloz, Academic Press, N. Y. (1969), p. 204 and in L. J.Haynes and F. H. Newth, Advances in Carbohydrate Chemistry, Vol. 10,Academic Press, 1955, pages 147-154. Other glycosyl halides which can beused in the subject invention are N-acetyl-2,3,4,7-tetra-O-acetyl-α andβ-lincosaminyl bromides as disclosed by B. Bannister, J. Chem. Soc.,Perkin, 3025 (1972). Still other substituted glycosyl halides which canbe used in the subject invention are3-acetamido-2,4,6-tri-O-benzyl-3-deoxyglucopyranosyl chloride [S. Kotoet al., Bull. Chem. Soc. Japan, 41, 2765 (1968)];2,3,4-tri-O-benzyl-6-(N-benzylacetamido)-6-deoxy-α-D-glucopyranosylchloride [Koto, ibid.];3-acetamido-2,4,6-tri-O-acetyl-3-deoxyglucopyranosyl bromide [Shibahara,et al., J. Amer. Chem. Soc., 94, 4353 (1972)]; and3,4,6-tri-O-acetyl-2-trifluoroacetamido-2-desoxy-α-D-glucopyranosylbromide [Meyer zv Reckendorf et al. Chem Ber., 103, 1792 (1970)].

Novel compounds produced by the novel process, hereinafter described indetail, can be shown generically by the formula ##STR2## wherein G is aglycosyl attached to the 3' or 6 position of paromamine, said glycosylof the formulae ##SPC11##

wherein R₁ is selected from the group consisting of OH, OAc, ##STR3##OCH₂ φ, NHR', NR' alkyl; wherein alkyl is from 1 to 5 carbon atoms,inclusive; R' is H or acyl of from 1 to 8 carbon atoms, inclusive; Ac isacetyl; and ##SPC12##

wherein R₂ is H or acetyl.

Further novel compounds produced by the novel process, hereinafterdescribed in detail, can be shown generically by the formula ##STR4##wherein A is a 4-O-(α-D-glycosyl)-2-deoxystreptamine selected from thegroup consisting of kanamine, gentamine C_(1a), gentamine C₁, gentamineC₂, gentamine C_(2b), gentamine B₁, nebramine, and lividamine, and G isglycosyl attached to the 6 position of said4-O-(α-D-glycosyl)-2-deoxystreptamine, said glycosyl of the formula##SPC13##

wherein R₁ is selected from the group consisting of OH, OAc, ##STR5##OCH₂ φ, NHR', NR' alkyl; wherein alkyl is from 1 to 5 carbon atoms,inclusive; R' is hydrogen or acyl of from 1 to 8 carbon atoms,inclusive; Ac is acetyl, ##SPC14##

wherein R₂ is H or acetyl, and ##SPC15##

wherein R₁ is as defined immediately above; excluding kanamycin A,kanamycin B, kanamycin C, nebramycin factor 4, nebramycin factor 5' andtobramycin.

The structural formulae for kanamycin A, kanamycin B, kanamycin C,nebramycin factor 4, nebramycin factor 5' and tobramycin are as follows:

    __________________________________________________________________________    Kanamycin A                                                                           R.sub.1 =NH.sub.2                                                                   R.sub.2 =OH                                                                            Nebramycin factor 4                                                                     R.sub.1 =OH                                                                         ##STR6##                               Kanamycin B                                                                           R.sub.1 =NH.sub.2                                                                   R.sub.2 =NH.sub.2                                                                      Nebramycin factor 5'                                                                    R.sub.1 =H                                                                          ##STR7##                               Kanamycin C                                                                           R.sub.1 =OH                                                                         R.sub.2 =NH.sub.2                                                                      Tobramycin                                                                              R.sub.1 =H                                                                         R.sub.2 =H                                6'                                                                           ##STR8##                                                                                             ##STR9##                                              __________________________________________________________________________

A sub-generic group of the novel compounds depicted supra can be shownas follows: ##STR10## wherein G is a glycosyl attached to the 3' or 6position of paromamine, said glycosyl of the formulae ##SPC16##

wherein R₁ is selected from the group consisting of OAc, OH, NHAc, orNH₂ ; and wherein Ac is acetyl.

Another sub-generic group of the novel compounds depicted supra can beshown as follows: ##STR11## wherein A is as defined above and G is aglycosyl attached to the 6 position of A, said glycosyl of the formulae##SPC17##

wherein R₁ is selected from the group consisting of OAc, OH, NHAc, orNH₂ ; and wherein Ac is acetyl; excluding kanamycin A, kanamycin B,kanamycin C, nebramycin factor 4, nebramycin factor 5' and tobramycin.

Another sub-generic group of the novel compounds depicted supra can beshown as follows: ##STR12## wherein G is a glycosyl attached to the 3'or 6 position of paromamine, said glycosyl of the formulae ##SPC18##

wherein R₃ is H or acetyl.

Another sub-generic group of the novel compounds depicted supra can beshown as follows: ##STR13## wherein A is as defined above and G is aglycosyl attached to the 6 position of A, said glycosyl of the formulae##SPC19##

wherein R₃ is H or acetyl.

The novel intermediates, disclosed herein, are useful to make novelaminoglycoside antibiotics. These novel aminoglycoside antibiotics areantibacterially active, and, thus, they can be used in variousenvironments to eradicate or control sensitive bacteria. Following arein vitro antibacterial test results for representative compounds of thesubject invention. The results were obtained with a standard disc plateassay using 12.5 mm paper discs.

    ______________________________________                                               Zone of Inhibition (mm)                                                 Compound                                                                              B. cereus            B. subtilis                                     Tested   5 mg/ml* 10 mg/ml*  5 mg/ml*                                                                             10 mg/ml*                                 ______________________________________                                        (7β)                                                                              25       34         35     38                                        (7α)                                                                             25       32         32     34                                        (9)      29       34         32     34                                        (11)     --       16         --     --                                        ______________________________________                                         *Concentration of Compound Tested.                                       

Compounds were also tested in a standard microplate test in Brain HeartInfusion (BHI) Agar, at a concentration of 1 mg/ml. Incubation is at 37°and end points are read at 20 hours. Brain Heart Infusion Agar (suppliedby Difco Laboratories, Detroit, Michigan, U.S.A.) has the followingcomposition:

    ______________________________________                                        Calf brains, infusion from                                                                            200 gm.                                               Beef heart, infusion from                                                                             250 gm.                                               Bacto Proteose-peptone, Difco                                                                         10 gm.                                                Bacto-Dextrose, Difco   1 gm.                                                 Sodium chloride         5 gm.                                                 Disodium phosphate      2.5 gm.                                               Agar                    15 gm.                                                ______________________________________                                    

    __________________________________________________________________________    Minimum Inhibitory Concentration (mcg/ml)                                     Compound      (9)  (7α)                                                                         Neamine Control                                                                         *(7β)                                                                         *Neamine Control                       __________________________________________________________________________    Organism                                                                      S. aureus 284 UC 76                                                                         125  1000 125     1000   62.5                                   S. aureus UC 570                                                                            250  500  250     1000   62.5                                   S. aureus UC 746                                                                            250  1000 250     500    62.5                                   S. hemolyticus UC 152                                                                       3.9  31.2 3.9     62.5   7.8                                    St. faecalis UC 694                                                                         >1000                                                                              >1000                                                                              1000    >1000  500                                    E. coli UC 45 250  500  62.5    500    125                                    P. vulgaris UC 93                                                                           500  1000 250     500    125                                    K. pneumoniae UC 58                                                                         62.5 62.5 7.8     62.5   15.6                                   S. schottmuelleri UC 126                                                                    250  500  62.5    250    62.5                                   Ps. aeruginosa UC 95                                                                        >1000                                                                              >1000                                                                              >1000   >1000  >1000                                  D. pneumoniae UC 41                                                                         >1000                                                                              >1000                                                                              >1000   >1000  >1000                                  __________________________________________________________________________     *Run on different day from other samples.                                     NOTE:                                                                         UC is a registered trademark designating The Upjohn Company Culture           Collection.                                                              

Compounds (7β), (7α) and (9) were tested again along with compounds (11)and (14β) against a neamine control using the same conditions as givenabove. These results are shown in the following table.

    __________________________________________________________________________    Minimum Inhibitory Concentration (mcg/ml)                                                               Neamine      Neamine                                Compound      (9) (11)                                                                              (7α)                                                                        Control                                                                            (7β)                                                                         (14β)                                                                        Control                                __________________________________________________________________________    Organism                                                                      S. aureus UC 76                                                                             125 1000                                                                              62.5                                                                              31.2 500 1000                                                                              31.2                                   S. hemolyticus UC 152                                                                       3.9 1000                                                                              7.8 2.0  250 1000                                                                              3.9                                    S. faecalis UC 694                                                                          1000                                                                              1000                                                                              1000                                                                              500  1000                                                                              1000                                                                              500                                    D. pneumoniae UC 41                                                                         125 1000                                                                              125 62.5 1000                                                                              1000                                                                              31.2                                   E. coli UC 45 250 1000                                                                              62.5                                                                              62.5 1000                                                                              1000                                                                              62.5                                   K. pneumoniae UC 58                                                                         31.2                                                                              1000                                                                              7.8 7.8  250 500 3.9                                    S. schottmuelleri UC 126                                                                    62.5                                                                              1000                                                                              125 62.5 500 1000                                                                              31.2                                   Ps. aeruginosa UC 95                                                                        1000                                                                              1000                                                                              1000                                                                              1000 1000                                                                              1000                                                                              1000                                   P. vulgaris UC 93                                                                           125 1000                                                                              125 15.6 1000                                                                              1000                                                                              62.5                                   P. mirabilis A-63                                                                           500 1000                                                                              500 250  1000                                                                              1000                                                                              250                                    P. morgani UC 3186                                                                          125 1000                                                                              125 31.2 1000                                                                              1000                                                                              62.5                                   P. rettgeri UC 339                                                                          1000                                                                              1000                                                                              1000                                                                              500  1000                                                                              1000                                                                              1000                                   S. marcescens UC 131                                                                        500 1000                                                                              250 125  1000                                                                              1000                                                                              250                                    S. flexneri UC 143                                                                          500 1000                                                                              125 62.5 1000                                                                              1000                                                                              62.5                                   S. typhi TG-3 250 1000                                                                              62.5                                                                              31.2 1000                                                                              1000                                                                              15.6                                   __________________________________________________________________________

Compound (14β) was also tested against a series of bacteria on astandard agar disc plate assay, as described above. These results are asfollows:

    ______________________________________                                        Agar Diffusion Assay of 3'-O-β-D-Ribosylneamine (14β)               ______________________________________                                                   B. cereus  B. subtilis S. aureus                                   Dilutions  UC 3145    UC 564      UC 76                                       Full Strength                                                                             25.5      37          25                                          1 : 2      24          35.5       23                                          1 : 4      21         33          21                                          1 : 8      18          30.5        17.5                                                  p. vulgaris                                                                              Ps. aeruginosa                                                                            E. coli                                     Dilutions  UC 93      UC 95       UC 51                                       Full Strength                                                                            26         20          27                                          1 : 2      22         17           24.5                                       1 : 4       19.5      14          22                                          1 : 8      16         --          18                                                     S. lutea   K. pneumoniae                                           Dilutions  UC 130     UC 57                                                   Full strength                                                                            25         31                                                      1 : 2       22.5      29                                                      1 : 4      19         27                                                      1 : 8      15         23                                                      ______________________________________                                    

From the above results, it is seen that compounds (7α), (7β), (9), (11)and (14β) are active against Bacillus cereus and, thus, these compoundscan be used to to treat woolen felts since B. cereus has been isolatedfrom deteriorated woolen felts in the paper industry. Compounds (7α),(7β), (9) and (14) are active against Bacillus subtilis, and, thus,these compounds can be used for controlling the infection of silkwormscaused by B. subtilis; further, these compounds can be used to minimizeor prevent odor in fish and fish crates caused by B. subtilis. Compounds(5α), (5β), (9) and (14β) are active against Staphylococcus aureus, and,thus, these compounds can be used as disinfectants on washed and stackedfood utensils contaminated with S. aureus. Compounds shown to be activeagainst Escherichia coli can be used to reduce, arrest, and eradicateslime production in papermill systems caused by this bacterium; theyalso can be used to prolong the life of cultures of Trichomonas foetus,Trichomonas hominis, and Trichomonas vaginalis by freeing them of E.coli contamination. Further, since some of the compounds are activeagainst Streptococcus hemolyticus, as shown above, they can be used todisinfect instruments, utensils, or surfaces, where the inactivation ofthis microorganism is desirable. Evidence of antibacterial activityagainst other bacteria, as shown above, is sufficient to enable theskilled artisan to use the compounds in a number of environments whichare well known to be inhabited by such bacteria.

Salts of the novel aminoglycoside antibiotics, disclosed herein, can bemade by reacting the parent antibiotic with a stoichiometric amount of anontoxic, pharmaceutically acceptable acid. Examples of such acids areacetic, hydrochloric, sulfuric, maleic, phosphoric, nitric, hydrobromic,ascorbic, malic, citric, and like acids used to make salts ofamine-containing antibiotics.

The following examples are illustrative of the process and products ofthe present invention but are not to be construed as limiting. Allpercentages are by weight and all solvent mixture proportions are byvolume unless otherwise noted. Temperatures are in centigrade.

The test data in the following preparation and Examples was obtained asfollows. Melting points were taken in a Thomas-Hoover m.p. apparatus.Infrared (ir) absorption spectra were recorded from mineral oil mulls ona Perkin-Elmer Infracord spectrophotometer. Proton magnetic resonance(pmr) spectra were recorded on a Varian A-60 spectrophotometer; allsamples were dissolved in deutero-acetone unless otherwise stated withtetramethylsilane as an internal standard. Chemical shifts are reportedas δ values (TMS 0.0). Optical rotations were observed at 25° in thesolvents noted at a concentration of 1%. Analtech (3 in. or 8 in.) tlcplates coated with silica gel G were used for tlc; visualization wasobtained by H₂ SO₄ charring unless stated otherwise. Tlc system J-18 isthe upper layer from equilibration of chloroform, methanol, concentratedNH₄ OH, water (25:25:3:47). Column chromatography used Silica Gel 60 (EMReagents, Elmsford, N.Y.). Fractions were monitored by tlc and combinedon the basis of the tlc profile.

Preparation of 2,3,5-Tri-O-acetyl-β-D-ribofuranosyl bromide ##SPC20##

Hydrogen bromide is passed for two minutes through a solution of 1.908 g(6 mmol) of 1,2,3,4-tetra-acetyl-β-D-ribofuranose at ambienttemperature. The solvent is distilled under vacuum. The residue isdissolved in 5 ml of toluene and then this solvent is removed at 1 mmolwith a bath temperature of 40°. This treatment is repeated. Tlc onsilica gel using chloroform-methanol (20:1) shows a very strong spot (H₂SO₄) at R_(f) 0.5 with a weak spot slower and a trace near the front.(1,2,3,4-Tetra-acetyl-β-D-ribofuranose) gives an R_(f) of 0.9 on thissystem. Pmr (CDCL₃) δ2.05-2.10 (3 singlets, COCH₃), 6.36 (s, anomeric αH), 6.7 (d, 4, anomeric βH) indicates a ratio of about 2:1 in favor of aβ-bromide (α-hydrogen) at C-1. The bromide is used without furtherpurification.

EXAMPLE 1 1,2',3,6'-Tetrakis-N-(trifluoroacetyl)neamine (2) ##SPC21##

Trifluoroacetic anhydride (33.5 ml, 160 mmol) is added at 15° ± 5° overa period of 30 minutes to a suspension of 9.66 g (30 mmol) of neamine in100 ml of acetonitrile and 22.4 ml (160 mmol) of triethylamine. Afterstirring at ambient temperature for 1 hour the solvent is evaporated invacuo. The residue is diluted with 150 ml of ethyl acetate. Theresulting solution is washed with 5% KHCO₃ -- saturated NaCl (1:1)several times, dried and concentrated. The residue is triturated withether and the crystals recrystallized from ethanol to give 15.95 g(75.3%) of (2), m.p. 286°-288° dec. An additional 1.2 g (5.2%) of (2),melting at 278°-280°, is obtained from the mother liquors. A portion isrecrystallized twice from ethanol to afford an analytical sample, m.p.304°-306°; [α]_(D) +63° (EtOH); ir 3600-3300 cm⁻ ¹ (NH/OH), 1700 (C =O), 1560 (amide II), 1220, 1180, 1160 (CF/C-O); pnr δ5.28 (d, 3,anomeric), 3.2-4.2 (cluster); mass spectrum (TMS deriv.) m/e 974 (M-15),497, 481.

Anal. Calcd. for C₂₀ H₂₂ F₁₂ N₄ O₁₀ : C, 34.00; H, 3.14; N, 7.93; F,32.28. Found: C, 33.76; H, 3.18; N, 8.12; F, 32.25.

EXAMPLE 2 5,6-O-Isopropylidene-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (3) and3',4',5,6-O-Diisopropylidene-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine(3a) ##SPC22##

A mixture of 7.06 g (10 mmol) of1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (2) in 30 ml ofacetonitrile and 60 ml of dimethoxypropane containing 0.25 ml oftrifluoroacetic acid is heated at reflux for 0.75 hour. Amberlite IRA-45(OH⁻) resin (12 g) supplied by Rohm and Haas Co., is added to the cooledsolution with stirring. After 10 minutes a neutral reaction is obtainedon moist acid-base indicator paper. The solution is filtered andconcentrated under vacuum. Chromatography over 500 g of silica gel usingchloroform methanol (10:1) for elution leads to the isolation of 5.68 g(76.25%) of noncrystalline monoketal (3). Rechromatography gives asample having the following data: [α]_(D) +75° (EtOH); ir 3430, 3300,3100 cm⁻ ¹ (NH/OH), 1705 (C = O), 1560 (amide II), 1215, 1185, 1160 (CF₃/c-O), pnr δ5.35 (d, 3, anomeric), 3.2-4.7 (cluster), 1.36 (S, >C(CH₃)₂.

Anal. Calcd. for C₂₃ H₂₆ F₁₂ N₄ O₁₀ : C, 37.00; H, 3.51; N, 7.51. Found:C, 36.72; H, 3.64; N, 7.58.

In addition to monoketal (3) a less polar fraction of 1.02 g (12.95%) isalso collected. Physical data indicates this to be diketal (3a): massspectrum m/e 771 (M--CH₃), 767 (M--F), 728 (M--(CH₃)₂ CO), 717 (M--CF₃),673 (M--CF₃ CONH₂), 377, 393; pmr δ5.48 (d, 3, anomeric), 3.2-4.5(cluster), 1.4 (S, 2 >C(CH₃)₂).

EXAMPLE 35,6-O-Isopropylidene-3',4'-bis-O-(p-nitrobenzoyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine(4) ##SPC23##

p-Nitrobenzoyl chloride (34.8 g, 0.19 mole) is added to a solution of36.0 g (0.048 mole) of ketal (3) in 420 ml of pyridine while cooling sothat the temperature remains below 35°. After stirring for 2.5 hours atambient temperature the pyridine is distilled under vacuum. The residueis dissolved in ethyl acetate and washed successively with dilute HCl,H₂ O and KHCO₃ solution. The residue after evaporation of the solventweighs 63.7 g. Chromatography over 5 kg of silica gel usingchloroformmethanol (40:1) for elution affords 47.5 g (94.2%) of solid(4); [α]_(D) -32° (acetone).

Anal. Calcd. for C₃₇ H₃₂ F₁₂ N₆ O₁₆ : C, 42.53; H, 3.09; N, 8.05. Found:C, 42.19; H, 3.01; N, 7.93.

EXAMPLE 43',4'-Bis-O-(p-nitrobenzoyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine(5) ##SPC24##

A solution of 45.5 g (43.6 mmol) of ketal (4) in 450 ml of 66% aceticacid solution is warmed at 65° for 4 hours. The reaction mixture islyophilized. The residue of 37.6 g is chromatographed over 3.5 kg ofsilica gel using chloroform-methanol (10:1) for elution. A fraction of35.4 g (80.7%) of diester (5) is obtained. It shows [α]_(D) acetone-43°; UV in EtOH λ max = 258 mμ (ε 26,700); ir in mineral oil mull, maxbands at 3250-3400 cm⁻ ¹ (NH/OH), 1705, 1750 (C = O), 1620 (C = C), 1575(amide II), 1220, 1180, 1160 (CF₃ /C:O); pmr δ 8.0-8.3 (aromatic), 5.64(d, 3, anomeric), 3.2-4.3 (cluster).

Anal. Calcd. for C₃₄ H₂₈ F₁₂ N₆ O₁₆ : C, 40.65; H, 2.81; N, 8.37. Found:C, 40.84; H, 2.96; N, 8.36.

EXAMPLE 53',4'-Bis-O-(p-nitrobenzoyl)-6-O-(2,3,5-tri-O-acetyl-α-D-ribofuranosyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (6α) and3',4'-bis-O-(p-nitrobenzoyl)-6-O-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine(6β) ##SPC25##

Benzene (50 ml) is distilled from a solution of 6.0 g (6 mmol) ofdiester (5) in 100 ml of purified nitromethane and 150 ml of benzene.2,3,5-O-Triacetyl-D-ribofuranosyl bromide (prepared as described abovefrom 11.8 mmol of tetraacetate) in 6 ml of nitromethane and 2.98 g (11.8mmol) of Hg(CN)₂ are added. Further additions of a total of 45.4 mmol ofbromide and 8.94 g of Hg(CN)₂ are made in two additions with about 2hours of reflux between additions. Tlc (chloroformmethanol, 20:1) showsthe absence of starting diester (5). Ethyl acetate (250 ml) is added andthe solution extracted twice with KHCO₃ solution. The dried solution isconcentrated under vacuum. Chromatography over 2 kg of silica gel usingchloroform-methanol (30-1) for elution followed by rechromatography offractions which are mixtures, gives 4.6 g (60.77%) of (6β) and 1.9 g(25%) of (6α). Rotations of -2° and -46° (acetone) are found for (6α)and (6β), respectively.

Anal. Calcd. for C₄₅ H₄₂ N₆ F₁₂ O₂₃ : C, 42.80; H, 3.35; N, 6.66. Found:(6α) C, 42.81; H, 3.53; N, 6.72; (6β) C, 42.85; H, 3.35; N, 6.55.

EXAMPLE 6 6-O-(β-D-Ribofuranosyl)neamine (7β) ##SPC26##

A solution of 2.17 g (1.72 mmol) of glycoside (6β) and 1.24 g (30.96mmol) of NaOH in 15 ml of methanol and 15 ml of water is heated atreflux for 15 minutes. The methanol is removed in vacuo. Water (75 ml)is added and the solution passed through 10 ml of Amberlite CG-50 (NH₄⁺) resin, supplied by Rohm and Haas Co. The column is washed with 100 mlH₂ O and then is eluted with a gradient of 400 ml each of water and 0.5NNH₄ OH. Fractions of about 40 ml are collected. The fractions aremonitored by dipped disc testing vs. B. cereus and tlc using systemJ-18. The results are summarized below.

    ______________________________________                                        Fraction No.                                                                             Zone Size (m mol)                                                                            Tlc Assay                                           ______________________________________                                        1-2        --             --                                                  3-5        --             (orange color, UV.sup.+)                            6-23       --             --                                                  24,25      tr             --                                                  26         23             1 spot                                              27         29             max. intensity                                      28         28             max. intensity                                      29         24             weaker                                              30         21             --                                                  31         20             --                                                  32         17             --                                                  33-35      --             --                                                  ______________________________________                                    

Fractions 26-29 are combined and lyophilized to give 460 mg (58.7%) of7β. Fractions 30-32 similarly yields 20 mg. Ir (Nujol) 3100-3500 cm⁻ ¹(NH/CH), 1600 (NH).

EXAMPLE 7 6-O-(α-D-Ribofuranosyl)neamine (7α) ##SPC27##

In the manner described for the β-isomer, 500 mg of glycoside recoveredfrom physical measurements is treated with 200 mg of NaOH and passedover CG-50 (NH₄ ⁺) to give 94 mg (53%) of (7α).

EXAMPLE 8 6-O-(dihydrogenorthoacetyl)-3',4'-bis-O-(p-nitrobenzoyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine,cyclic ester with 3,5-di-O-acetyl-α-D-ribofuranose (8) ##SPC28##

A solution of 3 g (3 mmol) of diol (5), 0.84 ml (6 mmol) oftriethylamine and 3 mmol of 2,3,5-tri-O-acetyl-D-ribofuranosyl bromidein 140 ml of tetrahydrofuran is heated at reflux. Tlc usingchloroform-methanol (20:1) indicates a new faster spot after 1 hour.Three additions of 3 mmol of bromide and 0.84 ml (6 mmol) oftriethylamine are made at hourly intervals. The reaction mixture isrefluxed an additional 4 hours, filtered and evaporated. The residue ischromatographed over 450 g of silica gel using chloroform-methanol(40-1) for elution. The product fraction weighs 2.10 g (55.6%). Diol (5)is recovered (977 mg) by stripping the column with chloroform-methanol,10:1. The orthoester (8) gives the following data: [α]_(D) -8°(acetone); ir, 3200-3600 cm⁻ ¹ (NH/OH), 1720-1770 (C = O), 1640 (C = C),1550 (amide II); pmr, δ (DMF) 7.7-8.2 -bis-(dimethylamino)-naphthalene(aromatic), 5.78 (d, 3, anomeric), 3.3-4.1 (cluster), 7.8 (S, COCH₃,CH₃) 7.4 (S, COCH₃).

Anal. Calcd. for C₄₅ H₄₂ F₁₂ N₆ O₂₃ : C, 42.80; H, 3.35; N, 6.66. Found:C, 42.41; H, 3.19; N, 6.34.

A similar product is obtained in 50% yield when 1,4-bis-(dimethylamino)is used instead of triethylamine.

EXAMPLE 9 6-O-Dihydrogen orthoacetylneamine, cyclic ester with3,5-di-O-acetyl-α-D-ribofuranose (9) ##SPC29##

Orthoester (8) (1.26 g, 1 mmol) in 9 ml of H₂ O and 9 ml MeOH containing720 mg (18 mmol) of NaOH is refluxed for 15 minutes. The methanol isevaporated. The aqueous residue is diluted with 50 ml H₂ O and passedthrough 60 ml of CG-50 (NH₄ ⁺). The column is eluted with a gradient of200 ml H₂ O and 200 ml of 0.5 N NH₄ OH. Fractions of 20 ml arecollected. They are assayed on tlc (J-18) and by dipped disc vs. B.cereus. Fractions 12-25 are combined on the basis of bioactivity and tlcdata and then lyophilized. There are thus obtained 406 mg of orthoester(9), ir (Nujol), 3100-3500 (NH/CH), 1600 (NH) (similar to neamine).

EXAMPLE 105,6-O-isopropylidene-3'-O-(2,3,4,6-tetrakis-O-acetyl-β-D-glucopyranosyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (10) and 5,6-O-Iisopropylidene3'-O-(2,3,4,6-tetrakis-O-acetyl-α-D-glucopyranosyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (10α) ##SPC30##

Twenty-five ml of benzene is distilled from a solution of 3.73 g (5mmol) of ketal (3) in 50 ml of purified CH₃ NO₂ and 75 ml of benzene.α-Acetobromoglucose (4.10 g, 10 mmol) and 2.5 g, 10 mmol of Hg(CN)₂ areadded and refluxed for 2 hours. Two more additions of bromide and baseare made at 2 hour intervals. The solvent is distilled under vacuum. Theresidue is dissolved in ethyl acetate, filtered, and washed with KHCO₃three times. Filtration is necessary to remove mercuric salts. Thesolution is dried and evaporated to give a residue of 14.3 g. Thismaterial is chromatographed over 0.75 kg of silica gel eluting withchloroform-methanol (20:1). A fraction of 5.03 g showing at least fourcomponents by tlc and a more polar fraction of 1.66 g (one component) isobtained. The latter shows in the ir spectrum bands at 1740 and 1550 cm⁻¹ indicative of ester and amide. On the bssis of the mobility and irdata it is characterized as glycoside (10).

EXAMPLE 113'-O-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine(10X) ##SPC31##

A solution of 1.6 g of glycoside (10) is dissolved in 30 ml of 66% HOAcand the mixture is heated at 65° for 2.5 hours. The solution islyophilized. Tlc (chloroform-methanol, 10:1) shows no (10) but a slowerspot (10X).

EXAMPLE 12 3'-O-(β-D-glucopyranosyl)neamine (11) ##SPC32##

Glycoside (10X) (1.11 g) and 720 mg of NaOH in 9 ml of MeOH and 9 ml ofH₂ O are refluxed for 15 minutes. The methanol is evaporated in vacuo.The aqueous solution is put over 50 ml of CG-50 (NH₄ ⁺). The column iseluted with 50 ml H₂ O and then with a gradient of 200 ml H₂ O and 200ml of 0.5N NH₄ OH. A fraction of 390 mg is obtained which seems to beone spot on silica gel tlc (J-18, ninhydrin).

using the dipped disc technique, glycoside (11) at 10 mg/ml, gives a 16mmol zone of inhibition vs. B. cereus. Neamine at 10 mg/ml gives a 32mmol zone and at 5 mg/ml a 29 mmol zone.

EXAMPLE 135,6-O-Isopropylidene-3'-O-(2,3,5,-tri-O-acetyl-62-D-ribofuranosyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (12β) ##SPC33##

A solution of 10 g of5,6-O-isopropylidene-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine (3)in 330 ml of benzene and 220 ml of nitromethane is heated to boiling and110 ml of distillate collected. 2,3,5-Tri-O-acetyl-β-D-ribofuranosylbromide is prepared from 16.5 g of1,2,3,5-tetra-O-acetyl-β-D-ribofuranose, as described above. The bromideis dissolved in 30 ml of nitromethane. One third of this solution and 7g of mercuric cyanide is added to the original reaction mixture. Themixture is heated to reflux. Two similar additions of bromide andmercuric cyanide are made after 1 and 2 hours of reflux. The mixture isrefluxed a final hour. The cooled reaction mixture is diluted with 750ml of ethyl acetate and extracted with two 500 ml portions of 5% KHCO₃solution. The organic layer is washed with brine and filtered throughNa₂ SO₄. The solvent is evaporated under vacuum. The residue ischromatographed over 1 kg of silica gel, and eluted withchloroform-methanol (20:1). Fractions of 50 ml are collected andmonitored by tlc. Fractions 1-30 contain fast moving impurities and arediscarded. Fractions 30-40 (27A) weigh 13.4 g, while fractions 41-48(27B) weigh 3.23 g. A 12.4 g portion of 27A is rechromatographed over 1kg of silica gel using chloroform-methanol (30:1) for elution. Thecenter cut based on tlc of the fractions is combined and evaporated togive 6.66 g of blocked analog (12β). CMR data, after removal of theblocking group as described infra, shows this compound to be chiefly the3' β isomer.

EXAMPLE 143'-O-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)-1,2',3,6'-tetrakis-N-(trifluoroacetyl)neamine(13β) ##SPC34##

A solution of 6.66 g of (12β) in 40 ml of acetic acid and 20 ml of wateris heated at reflux for 2 hours. The solvent is removed bylyophilization. The residue of 5.038 g is used in the next step, Example15, without purification.

EXAMPLE 15 3'-O-(β-D-Ribofuranosyl)neamine (14β) ##SPC35##

The residue from Example 14 is dissolved in 73 ml of methanol-water(1:1) containing 2.93 g of NaOH. The solution is refluxed for 0.5 hour,cooled and 12 ml of N HCl is added. The methanol is evaporated undervacuum. The aqueous residue is diluted with 200 ml of water and passedthrough 250 ml of Amberlite CG-50 (NH₄ ⁺ form). The column is washedwith 250 ml H₂ O and then eluted with a gradient composed of 1 l ofwater and 1 l of 0.5N NH₄ OH. Fractions (50 ml) are monitored byinhibition of growth of B. cereususing the dipped disc technique andalso by tlc (J-18 system) visualizing with ninhydrin. Fractions having azone of inhibition of greater than 18 mmol and also showing a goodresponse on tlc are combined and lyophilized. A yield of 1.47 g of whitepowder is obtained. CMR data indicates this to be chiefly 14β.

EXAMPLE 16

By substituting the 2,3,5-O-triacetyl-D-ribofuranosyl bromide in Example5 by:

2,3,4,6 Tetra-O-acetyl-β-D-altropyranosyl chloride

2,3,4-Tri-O-acetyl-β-L-arabinopyranosyl chloride

3,4-Di-O-acetyl-2-deoxy-D-ribopyranosyl chloride

2,3,4,6-Tetra-O-acetyl-α-D-galactopyranosyl chloride

2,3,4,6 Tetra-O-acetyl-β-D-galactopyranosyl chloride

2,3,4,6 -Tetra-O-acetyl-α-D-glucopyranosyl chloride

2,3,4,6-Tetra-O-benzoyl-α-D-glucopyranosyl chloride

2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl chloride

2,3,4,6-Tetra-O-acetyl-α-D-mannopyranosyl chloride

2,3,4,6-Tetra-O-benzoyl-α-D-mannopyranosyl chloride

2,3,4-Tri-O-acetyl-α-L-rhamnopyranosyl chloride

2,3,4-Tri-O-benzoyl-α-L-rhamnopyranosyl chloride

2,3,5-Tri-O-acetyl-α-D-ribofuranosyl chloride

2,3,4-Tri-O-benzoyl-α-D-ribopyranosyl chloride

2,3,4-Tri-O-acetyl-β-D-ribopyranosyl chloride

2,3,4-Tri-O-benzoyl-β-D-ribopyranosyl chloride

2,3,4-Tri-O-acetyl-α-D-xylopyranosyl chloride

2,3,4-Tri-O-acetyl-β-D-xylopyranosyl chloride

2,3,4-Tri-O-acetyl-6-deoxy-α-D-glucopyranosyl chloride

2,3,4-Tri-O-acetyl-β-D-arabinopyranosyl bromide

2,3,4-Tri-O-benzoyl-β-D-arabinopyranosyl bromide

3,4,6-Tri-O-acetyl-2-deoxy-α-D-glucopyranosyl bromide

3,4,6-Tri-O-benzoyl-2-deoxy-α-D-glucopyranosyl bromide

2,3,4-Tri-O-acetyl-6-deoxy-α-D-glucopyranosyl bromide

1,3,4,5-Tetra-O-acetyl-β-D-fructopyranosyl bromide

1,3,4,5-Tetra-O-benzoyl-β-D-fructopyranosyl bromide

2,3,4,6-Tetra-O-acetyl-α-D-galactopyranosyl bromide

2,3,4,6-Tetra-O-acetyl-α-D-glucopyranosyl bromide

2,3,4-Tri-O-acetyl-6-O-benzoyl-α-D-glucopyranosyl bromide

2,3,4-Tri-O-acetyl-6-O-methyl-α-D-glucopyranosyl bromide

6-O-Acetyl-2,3,4-Tri-O-benzyl-α-D-glucopyranosyl bromide

2,3,4,6-Tetra-O-acetyl-α-D-mannopyranosyl bromide

2,3,4,6-Tetra-O-benzoyl-α-D-mannopyranosyl bromide

2,3,4-Tri-O-acetyl-α=L-rhamnopyranosyl bromide

2,3,4-Tri-O-benzoyl-α-L-rhamnopyranosyl bromide

2,3,4-Tri-O-acetyl-β-D-ribopyranosyl bromide

2,3,4-Tri-O-benzoyl-β-D-ribopyranosyl bromide

2,3,4-Tri-O-benzoyl-D-xylopyranosyl bromide

2,3,4-Tri-O-acetyl-L-xylopyranoxyl bromide

2,3,4-Tri-O-benzoyl-L-xylopyranoxyl bromide

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl bromide

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-α-D-glucopyranosyl chloride

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl chloride

2-Benzamido-3,4,6-tri-O-benzoyl-2-deoxy-α-D-glucopyranosyl bromide

3,4,6-Tri-O-acetyl-2-benzamido 2-deoxy-α-D-glucopyranosyl chloride

3,4,6-Tri-O-acetyl-2-[(benzyloxycarbonyl)-amino]-2-deoxy-α-D-glucopyranosylbromide

3,4,6-Tri-O-acetyl-2-deoxy-2-(2,4-dinitroanilino)-α-D-glucopyranosylbromide

2-Acetamido-3,4-di-O-acetyl-2-deoxy-D-ribofuranosyl chloride

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-D-galactopyranosyl bromide

2-Acetamido-3,4,6-tri-O-acetyl-2-deoxy-α-D-galactopyranosyl chloride

3-Acetamido-2,4,6-tri-O-acetyl-3-deoxy-α-D-mannopyranosyl bromide

3-Acetamido-2,4,6-tri-O-acetyl-3-deoxy-α-D-mannopyranosyl chloride

2,4,6-Tri-O-acetyl-3-[(benzyloxycarbonyl)-amino]-3-deoxy-α-D-glucopyranosylbromide

2,3,4-Tri-O-acetyl-6-[(benzyloxycarbonyl)-amino]-6-deoxy-α-D-glucopyranosylbromide

2,4,6-Tri-O-acetyl-3-[(benzyloxycarbonyl)-amino]-3-deoxy-D-glucopyranosylchloride

2,3,4-Tri-O-acetyl-6-[(benzyloxycarbonyl)-amino]-6-deoxy-D-glucopyranosylchloride

N-acetyl-2,3,4,7-tetra-O-acetyl-α and β-1 incosaminyl bromides

3-Acetamido-2,4,6-tri-O-benzyl-3-deoxyglucopyranosyl chloride

2,3,4-Tri-O-benzyl-6(N-benzylacetamido)-6-deoxy-α-D-glucopyranosylchloride

3-Acetamido-2,4,6-tri-O-acetyl-3-deoxyglucopyranosyl bromide

3,4,6-Tri-O-acetyl-2-trifluoroacetamido-2-desoxy-α-D-glucopyranosylbromide

there are obtained the corresponding 6 -O-glycosyl analogs of neaminehaving ester and amino protecting groups. These protecting groups arethen removed by following the procedure of Example 6 to affordantibacterially active 6 -O-glycosyl analogs of neamine.

EXAMPLE 17

By substituting the α-acetobromoglucose in Example 10 by 2,3,5-O-triacetyl=D-ribofuranosyl bromide there is obtained the corresponding3'-O-D-glycosyl analog of neamine having ester and amino protectinggroups. These protecting groups are then removed by following theprocedure of Examples 11 and 12 to afford antibacterially active3'-O-D-glycosyl analogs of neamine.

EXAMPLE 18

By substituting the α-acetobromoglucose in Example 10 by the glycosylhalides in Example 16, there are obtained the corresponding 3'-O-D-glycosyl analogs of neamine having ester and amino protectinggroups. These protecting groups are then removed by following theprocedure of Examples 11 and 12 to afford antibacterially active 3'-O-D-glycosyl analogs of neamine.

EXAMPLE 19

1-N-AHBA derivative of the 6 -O- and 3'-O-D-glycosyl analogs of neamine,as prepared in the preceding examples, is made by first blocking the6'-amino of the amino-glycoside by reacting it withN-benzyloxycarbonyloxylsuccinimide in aqueous dimethylformamide to formthe 6'-N-carbenzoxyamino glycoside. This compound is then selectively1-N-acylated with L(-)γ-benzyloxycarbonylamino-α-hydroxybutyric acid,N-hydroxysuccinimide ester in aqueous ethylene glycol dimethyl ether.The carbobenzoxy groups at 6'-N and at the γ-N are then removed byhydrogenolysis using palladium on charcoal as catalyst.

EXAMPLE 20

By substituting the 2,3,5 -tri-O-acetyl-D-ribofuranosyl bromide inExample 8 by the glycosyl halides in Example 16, there are obtained thecorresponding 6-O-D-glycosyl ortho esters of neamine having ester andamino protecting groups. These protecting groups are then removed byfollowing the procedure of Example 9 to afford antibacterially active 6-O-D-glycosyl ortho esters of neamine.

EXAMPLE 21

By substituting compound (5) in Example 8 by compound (3), there isobtained the cyclic ester with 3,5 -di-O-acetyl-α -D-ribofuranose of the5,6-ketal compound (3). The ketal moiety is removed by following theprocedures of Example 4, and the ester and amino protecting groups areremoved by following the procedures of Example 9 to affordantibacterially active 3' -O-D ortho esters of neamine.

EXAMPLE 22

By substituting the 2,3,5 -tri-O-acetyl-D-ribofuranosyl bromide inExample 8 by the glycosyl halides in Example 16, and compound (5) inExample 8 by compound (3), there are obtained the corresponding esterand amino protected ortho esters of the 5,6-ketal compound (3). Theketal moiety, and the ester and amino protecting groups are removed bythe procedures referred to in Example 21 to afford correspondingantibacterially active 3'-O-D-glycosyl ortho esters of neamine.

Preparation of Neamine

Neamine can be prepared from neomycin B by the procedures disclosed inU.S. Pat. No. 2,691,675. It also can be synthesized from paromamine asdisclosed by S. Umezawa and K. Tatsuta in Bull. Chem. Soc. Japan, 402371-75 (1967).

I claim:
 1. A compound of the formula ##EQU1## wherein G is a glycosylattached to the 3' or 6 position of paromamine, said glycosyl of theformulae ##SPC36##wherein R₁ is selected from the group consisting ofOH, OAc, ##EQU2## OCH₂ φ, NHR', NR' alkyl; wherein alkyl is from 1 to 5carbon atoms, inclusive; R' is H or acyl of from 1 to 8 carbon atoms,inclusive; Ac is acetyl; and ##SPC37## wherein R₂ is H or acetyl; andnontoxic pharmaceutically acceptable acid addition salts thereof.
 2. Acompound of the formula, according to claim 1 ##STR14## wherein G is aglycosyl attached to the 3' or 6 position of paromamine, said glycosylof the formulae ##SPC38##wherein R₁ is selected from the groupconsisting of OAc, OH, NNAc, or NH₂ ; and wherein Ac is acetyl; andnontoxic pharmaceutically acceptable acid addition salts thereof.
 3. Acompound of the formula, according to claim 2 ##STR15## wherein G is aglycosyl attached to the 3' or 6 position of paromamine, said glycosylof the formulae ##SPC39##wherein R₃ is H or acetyl; and nontoxicpharmaceutically acceptable acid addition salts thereof.
 4. A compoundof the formula ##STR16## wherein A is a 4-O-(α-D-glycosyl)-2-deoxystreptamine selected from the group consisting ofkanamine, gentamine C_(1a), gentamine C₁, gentamine C₂, gentamineC_(2b), gentamine B₁, nebramine, and lividamine, and G is a glycosylattached to the 6 position of said 4-O-(α-D-glycosyl)-2-deoxystreptamine, said glycosyl of the formula##SPC40##wherein R₁ is selected from the group consisting of OH, OAc,##STR17## OCH₂ φ, NHR', NR' alkyl; wherein alkyl is from 1 to 5 carbonatoms, inclusive; R' is hydrogen or acyl of from 1 to 8 carbon atoms,inclusive; Ac is acetyl, ##SPC41## wherein R₂ is H or acetyl, andnontoxic pharmaceutically acceptable acid addition salts thereof.
 5. Acompound of the formula, according to claim 4 ##STR18## wherein A is asdefined in claim 4 and G is a glycosyl attached to the 6 position of A,said glycosyl of the formula ##SPC42##wherein R₁ is selected from thegroup consisting of OAc, OH, NHAc, or NH₂ ; and wherein Ac is acetyl;and nontoxic pharmaceutically
 6. A compound of the formula, according toclaim 5 ##STR19## wherein A is as defined in claim 4 and G is a glycosylattached to the 6 position of A, said glycosyl of the formula##SPC43##wherein R₃ is H or acetyl; and nontoxic pharmaceuticallyacceptable acid addition salts thereof.
 7. A compound of the formula##STR20## wherein n is an integer of from 0 to 2, inclusive; and G is asdefined in claim
 1. 8. A compound of the formula ##STR21## wherein n isan integer of from 0 to 2, inclusive; and G is as defined in claim
 2. 9.A compound of the formula ##STR22## wherein n is an integer of from 0 to2, inclusive; and G is as defined in claim
 3. 10. A compound of theformula ##STR23## wherein n is an integer of from 0 to 2, inclusive; andA and G are as defined in claim
 4. 11. A compound of the formula##STR24## wherein n is an integer of from 0 to 2, inclusive; and A and Gare as defined in claim
 5. 12. A compound of the formula ##STR25##wherein n is an integer of from 0 to 2, inclusive; and A and G are asdefined in claim
 6. 13. A process for preparing a compound, as definedin claim 4, which comprises:1. selectively blocking the amino groups onthe aglycone,
 2. selectively blocking the 5,6-hydroxyls of theamino-protected aglycone to form a ketal,
 3. selectively acylating the3',4'-hydroxyls of the amino-protected aglycone wherein the5,6-hydroxyls are blocked as the ketal,
 4. selectively removing saidketal group to form an amino-protected aglycone having the3'-4'-hydroxyls acylated,
 5. subjecting said acylated amino-protectedaglycone to a glycosylation reaction with a glycosyl halide selectedfrom the group consisting of ##SPC44## wherein Z is Cl or Br; R₁ = OAc,##STR26## OCH₂ φ, NHR', NR' alkyl; alkyl = 1-5 carbon atoms, inclusive:R' = acyl of from 1 to 8 carbon atoms, inclusive; Ac = acetyl; and##SPC45## wherein Ac is acetyl, and
 6. selectively removing theamino-protecting groups and the acyl moieties to afford the compound, asdefined in claim
 4. 14. A process, as defined in claim 13, wherein theaglycone amino groups are selectively blocked as the trifluoroacetates.15. A process, according to claim 13, wherein the said ketal is formedby reacting the amino-protected aglycone in acetonitrile and a dialkoxylower alkane wherein the alkoxy and lower alkane are from 1 to 8 carbonatoms, inclusive, in the presence of an acid catalyst.
 16. A process,according to claim 13, wherein the said selective acylation of the 3'and 4' hydroxyls is conducted with an acylating agent selected from thegroup consisting of a halide or anhydride of a hydrocarbon carboxylicacid of from 2 to 18 carbon atoms, inclusive; or a halo-, nitro-,hydroxy-, amino-, cyano-, thiocyano-, and lower alkoxy- substitutedhydrocarbon carboxylic acid of from 2 to 18 carbon atoms, inclusive, inthe presence of an acid-binding agent.
 17. A process, according to claim13, wherein the said 5,6-ketal group is selectively removed by mild acidhydrolysis.
 18. A process, according to claim 13, wherein the saidglycosylation is conducted under anhydrous conditions using an excess ofthe glycosyl halide in nitromethanebenzene in the presence of mercuriccyanide.
 19. A process, according to claim 13, wherein the said esterand amino-protecting groups are selectively and simultaneously removedwith a strong aqueous alkali of about 2N in methanol.
 20. A process forpreparing the compound of claim 1, wherein the glycosyl is on the 3'position of paromamine, which comprises:1. selectively blocking theamino groups on paromamine,
 2. selectively blocking the 5,6-hydroxyls ofthe amino-protected paromamine to form a ketal,
 3. subjecting saidamino-protected paromamine 5,6-ketal to a glycosylation reaction with aglycosyl halide selected from the group consisting of ##SPC46## whereinZ is Cl or Br; R₁ = OAc, ##STR27## OCH₂ O, NHR', NR' alkyl; alkyl = 1-5carbon atoms, inclusive; R' = acyl of from 1 to 8 carbon atoms,inclusive; Ac = acetyl; and ##SPC47## wherein Ac is acetyl, 4.selectively removing the 5,6-ketal moiety, and5. selectively removingthe amino-protecting groups to afford the compound, as defined inclaim
 1. 21. A process, according to claim 20, wherein the paramamineamino groups are selectively blocked as the trifluoroacetates.
 22. Aprocess, according to claim 20, wherein the said ketal is formed byreacting the amino-protected paromamine in acetonitrile and a dialkoxylower alkane wherein the alkoxy and lower alkane are from 1 to 8 carbonatoms, inclusive, in the presence of an acid catalyst.
 23. A process,according to claim 20, wherein the said glycosylation is conducted underanhydrous conditions using an excess of the glycosyl halide innitromethanebenzene in the presence of mercuric cyanide.
 24. A process,according to claim 20, wherein the said 5,6-ketal group is selectivelyremoved by mild acid hydrolysis.
 25. A process, according to claim 20,wherein the said amino-protecting groups are selectively removed with astrong aqueous alkali of about 2N in methanol.
 26. A process forpreparing a compound, as defined in claim 1, wherein the glycosyl is onthe 6 position of paromamine, which comprises:1. selectively blockingthe amino groups on paromamine
 2. selectively blocking the 5,6-hydroxylsof the amino-protected paromamine to form a ketal,
 3. selectivelyacylating the 3' ,4'-hydroxyls of the amino-protected paromamine whereinthe 5,6-hydroxyls are blocked as the ketal,
 4. selectively removing saidketal group to form an amino-protected paromamine having the3'-4'-hydroxyls acylated,
 5. subjecting said acylated amino-protectedparomamine to a glycosylation reaction with a glycosyl halide selectedfrom the group consisting of ##SPC48## wherein Z is Cl or Br; R₁ = OAc,##STR28## OCH₂ φ, NHR', NR' alkyl; alkyl = 1-5 carbon atoms, inclusive;R' = acyl of from 1 to 8 carbon atoms, inclusive; Ac = acetyl; and##SPC49## wherein Ac is acetyl, and
 6. selectively removing theamino-protecting groups and the acyl moieties to afford the compound, asdefined in claim
 1. 27. A process, as defined in claim 26, wherein theparomamine amino groups are selectively blocked as thetrifluoroacetates.
 28. A process, according to claim 26, wherein saidketal is formed by reacting the amino-protected paromamine inacetonitrile and a dialkoxy lower alkane wherein the alkoxy and loweralkane are from 1 to 8 carbon atoms, inclusive, in the presence of anacid catalyst.
 29. A process, according to claim 26, wherein the saidselective acylation of the 3' and 4' hydroxyls is conducted with anacylating agent selected from the group consisting of a halide oranhydride of a hydrocarbon carboxylic acid of from 2 to 18 carbon atoms,inclusive; or a halo-, nitro-, hydroxy-, amino-, cyano-, thiocyano-, andlower alkoxy-substituted hydrocarbon carboxylic acid of from 2 to 18carbon atoms, inclusive, in the presence of an acid-binding agent.
 30. Aprocess, according to claim 26, wherein the said 5,6-ketal group isselectively removed by mild acid hydrolysis.
 31. A process, according toclaim 26, wherein the said glycosylation is conducted under anhydrousconditions using an excess of the glycosyl halide in nitromethanebenzenein the presence of mercuric cyanide.
 32. A process, according to claim26, wherein the said ester and amino-protecting groups are selectivelyand simultaneously removed with a strong aqueous alkali of about 2N inmethanol.